Alkaline Phasphatase
conjugation
Known as alkaline phosphatase labeling, is a common method used in biochemistry and molecular biology to label proteins, antibodies, or other molecules with alkaline phosphatase enzymes. These conjugates are widely used in various applications, including enzyme-linked immunosorbent assays (ELISAs), immunohistochemistry (IHC), western blotting, and in situ hybridization
Multiplexing
Alkaline phosphatase conjugates can be combined with other enzyme conjugates or detection methods to enable multiplexed detection of multiple target molecules within the same sample.
Versatility
Alkaline phosphatase conjugates can be used for the detection of various target molecules, including proteins, peptides, antibodies, nucleic acids, and small molecules. They are compatible with different assay formats and detection methods.
Stability
Alkaline phosphatase conjugates are stable under a wide range of pH and temperature conditions, making them suitable for long-term storage and various experimental conditions.
Alkaline Phasphatase Conjugation Applications
Enzyme-Linked Immunosorbent Assay (ELISA)
AP-conjugated secondary antibodies are widely used in ELISA for the detection of target antigens. The enzymatic activity of AP catalyzes the conversion of colorimetric or chemiluminescent substrates, allowing for sensitive and quantitative detection of antigen-antibody interactions.
Immunohistochemistry (IHC) and Immunofluorescence (IF)
AP-conjugated secondary antibodies are used in IHC and IF assays for the detection of specific antigens in tissue sections or cultured cells. AP-conjugated antibodies can generate permanent staining in IHC or fluorescent signals in IF, enabling the visualization and localization of target proteins.
Western Blotting
AP-conjugated secondary antibodies are used in western blotting for the detection of target proteins separated by gel electrophoresis. AP catalyzes the conversion of chromogenic or chemiluminescent substrates, allowing for the visualization and quantification of protein bands on the blot.
In Situ Hybridization (ISH)
AP-conjugated oligonucleotide probes are used in ISH assays for the detection of specific nucleic acid sequences in tissue sections or whole-mount preparations. AP catalyzes the conversion of chromogenic or chemiluminescent substrates, enabling the visualization of mRNA or DNA expression patterns.
Flow Cytometry
AP-conjugated antibodies or ligands can be used in flow cytometry assays for the detection of cell surface markers or intracellular proteins. AP-conjugated probes provide an alternative detection method in flow cytometry, allowing for the simultaneous analysis of multiple parameters.
Multiplexed Assays
AP conjugation can be combined with other enzyme conjugates, such as horseradish peroxidase (HRP) or β-galactosidase, to enable multiplexed detection of multiple targets within the same sample. This allows for the simultaneous analysis of different analytes in complex biological samples.
Diagnostic Assays
AP-conjugated antibodies or ligands are used in diagnostic assays for the detection of various biomarkers, pathogens, or disease-associated molecules. AP-based assays are employed in clinical laboratories for the diagnosis and monitoring of infectious diseases, autoimmune disorders, and cancer biomarkers.
Overall, alkaline phosphatase conjugation offers a reliable and versatile approach for the detection and quantification of target molecules in a wide range of biological samples, making it a valuable tool in research, diagnostics, and biotechnology applications.